title: Utility of specimens positive for Neisseria gonorrhoeae by the Aptima Combo 2 assay for assessment of strain diversity and antibiotic resistance.
abstract: In our jurisdiction, the Aptima Combo 2 assay (Gen-Probe, Inc.) is used to detect Neisseria gonorrhoeae from specimens collected at clinics for sexually transmitted infections (STI) and from select community patients. In addition, swabs are also collected for N. gonorrhoeae culture, susceptibility testing, and sequence typing (ST). Since only a small proportion of samples from provincial cases undergo culture, the available trends in antimicrobial susceptibility and predominant strain types may not be representative of all N. gonorrhoeae infections. Due to the limitations facing the use of N. gonorrhoeae culture to understand these trends in the general community, we performed a molecular analysis for markers of cephalosporin resistance and ST determination by using nucleic acid extracts of specimens sent for Aptima testing. Thirty-four samples submitted for both Aptima testing and N. gonorrhoeae culture from the same anatomic location (within 24 h) were included in the study. Sequence type was determined based on the sequence of the por and tbpB genes, and amino acid changes in the PBP 2 protein, encoded by the penA gene, were considered representative for the assessment of antimicrobial susceptibility. Sequence identity of 100% was observed between the sequences obtained from Aptima-analyzed samples and culture samples. Sequencing results showed an association between decreased susceptibility to extended-spectrum cephalosporins (ESC(ds)), tbp allele 110, ST 1407, and amino acid changes (G545S, I312M, and V316T) in the PBP 2 protein. Our data, generated based on a few representative genes, suggest that gonococcal samples positive by Aptima testing can be used to determine single nucleotide polymorphisms associated with ESC(ds) and the sequence type based on molecular strain typing. Confirmation of these findings may obviate the need for gonorrhea culture in the future.
authors.affiliation: Provincial Laboratory for Public Health, Calgary, Alberta, Canada.
authors.forename: Kanti
authors.initials: K
authors.lastname: Pabbaraju
authors.forename: Sallene
authors.initials: S
authors.lastname: Wong
authors.forename: Joanna J
authors.initials: JJ
authors.lastname: Song
authors.forename: Ameeta E
authors.initials: AE
authors.lastname: Singh
authors.forename: Ron
authors.initials: R
authors.lastname: Read
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Drews
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001431:Bacteriological Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D024881:Drug Resistance, Bacterial; D005260:Female; D005798:Genes, Bacterial; D006069:Gonorrhea / Q000382:microbiology; D006801:Humans; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D020125:Mutation, Missense; D009344:Neisseria gonorrhoeae / Q000145:classification* / Q000187:drug effects* / Q000302:isolation & purification; D020641:Polymorphism, Single Nucleotide
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial